Article

Role of the bilayer in the shape of the isolated erythrocyte membrane.

Biophysics Section Boston University School of Medicine 02118 Boston Massachusetts; Department of Biochemistry University of Chicago 60637 Chicago Illinois
Journal of Membrane Biology (Impact Factor: 2.48). 02/1982; 69(2):113-23. DOI: 10.1007/BF01872271
Source: PubMed

ABSTRACT The determinants of cell shape were explored in a study of the crenation (spiculation) of the isolated erythrocyte membrane. Standard ghosts prepared in 5 mM NaPi (pH 8) were plump, dimpled disks even when prepared from echinocytic (spiculated) red cells. These ghosts became crenated in the presence of isotonic saline, millimolar levels of divalent cations, 1 mM 2,4-dinitrophenol or 0.1 mM lysolecithin. Crenation was suppressed in ghosts generated under conditions of minimal osmotic stress, in ghosts from red cells partially depleted of cholesterol, and, paradoxically, in ghosts from red cells crenated by lysolecithin. The susceptibility of ghosts to crenation was lost with time; this process was potentiated by elevated temperature, low ionic strength, and traces of detergents or chlorpromazine. In that ghost shape was influenced by a variety of amphipaths, our results favor the premise that the bilayer and not the subjacent protein reticulum drives ghost crenation. The data also suggest that vigorous osmotic hemolysis induces a redistribution of lipids between the two leaflets of the bilayer which affects membrane contour through a bilayer couple mechanism. Subsequent relaxation of that metastable distribution could account for the observed loss of crenatability.

0 Bookmarks
 · 
44 Views
  • [Show abstract] [Hide abstract]
    ABSTRACT: Lipid rafts on cell membranes have heterogeneity such as cholesterol-rich microdomains and sphingolipids-rich microdomains. We previously reported that beta-cyclodextrin (beta-CyD) induced morphological changes of red blood cells (RBC) from discocyte to stomatocyte, possibly due to extraction of cholesterol from cholesterol-rich lipid rafts of RBC membranes. In this study, the effects of methyl-beta-cyclodextrin (M-beta-CyD) and 2,6-di-O-methyl-beta-cyclodextrin (DM-beta-CyD) on lipid rafts and morphological changes in rabbit RBC (RRBC) were examined, compared to those of beta-CyD. In sharp contrast to beta-CyD, M-beta-CyD and DM-beta-CyD induced morphological changes of RRBC from discocyte to echinocyte. At pre-hemolytic concentrations of beta-CyDs, M-beta-CyD and DM-beta-CyD strongly released cholesterol from cholesterol-rich lipid rafts, compared to beta-CyD. Meanwhile, the lowering effects of DM-beta-CyD on fluorescent sphingomyelin analogue in sphingolipids-rich lipid rafts were more potent than those of beta-CyD and M-beta-CyD. The magnitude of the abilities of M-beta-CyD and DM-beta-CyD to extract membrane constituents was higher than that of beta-CyD, consistent with that of hemolytic activity. Furthermore, DM-beta-CyD and M-beta-CyD, not beta-CyD, lowered the amount of proteins in cholesterol-rich lipid rafts of RRBC. These results suggest that higher hemolytic activity and morphological changes from discocyte to echinocyte in RRBC induced by M-beta-CyD and DM-beta-CyD may be due to the extraction of both cholesterol and proteins from cholesterol-rich lipid rafts of RRBC, although DM-beta-CyD may interact with sphingolipids-rich lipid rafts on RRBC membranes only slightly.
    Biological & Pharmaceutical Bulletin 05/2009; 32(4):700-5. · 1.85 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Morphological response (MR) of red blood cells represents a triphasic sequence of spontaneously occurring shape transformation between different shape states upon transfer the cells into isotonic sucrose solution in the order: S(0) (initial discoid shape in physiological saline)-->S(1) (echinocytic shape at the beginning of MR, phase 1)-->S(2) (intermediate discoid shape, phase 2)-->S(3) (final stomatocytic shape, phase 3). In this paper, the dynamics of cell shape changes was investigated by non-invasive light fluctuation method and optical microscopy. Among 12 possible transitions between four main shape states, we experimentally demonstrate here an existence of nine transitions between neighbour or remote states in this sequence. Based on these findings and data from the literature, we may conclude that red blood cells are able to change their shape through direct transitions between four main states except transition S(1)-->S(0), which has not been identified yet. Some shape transitions and their temporal sequence are in accord with predictions of bilayer couple concept, whereas others for example transitions between remote states S(3)-->S(1), S(1)-->S(3) and S(3)-->S(0) are difficult to explain based solely on the difference in relative surface areas of both leaflets of membrane suggesting more complex mechanisms involved. Our data show that MR could represents a phenomenon in which the major role can play pH and chloride-sensitive sensor and switching mechanisms coupled with transmembrane signaling thus involving both cytoskeleton and membrane in coordinated shape response on changes in cell ionic environment.
    Biochimica et Biophysica Acta 09/2010; 1798(9):1767-78. · 4.66 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: The aim of this study is to analyze the electromagnetic energy stored in stomatocyte, erythrocyte and echinocyte cells exposed to a linearly polarized electromagnetic plane wave at 900, 1800 and 2450MHz radiofrequency signals. This analysis can provide a better understanding of the order of appearance of altered shapes of erythrocytes (RBC) in the stomatocyte-echinocyte transition under radiofrequency exposure in terms of the deposited electromagnetic energy. For this purpose we use a realistic geometrical cell model based on parametric equations that allow for continuous transformations between normal erythrocytes and three stomatocyte subclasses with different degree of invagination and also between normal erythrocytes and echinocytes with an arbitrary number of spicules. We use a finite element technique with adaptive meshing for calculating the electromagnetic energy deposited on the different regions of the cell models. It is found that the echinocyte cell stores the minimum electromagnetic energy and therefore from an energetic point of view it would be the most stable and preferred cell state when this electromagnetic energy is the predominant energy component.
    Bioelectrochemistry (Amsterdam, Netherlands) 08/2009; 77(2):158-61. · 2.65 Impact Factor